High Potential for Anaerobic Microbial Sulfur Oxidation in Oil Sands Tailings Ponds

The biogenic production of toxic H2S gas in sulfate-rich oil sands tailings ponds is associated with strong environmental concerns. Beside precipitation into sulfide minerals and chemical re-oxidation, microbial sulfur oxidation may catalyze sulfide re-cycling but potentially contributes to acid rock drainage (ARD) generation. To evaluate the microbial potential for sulfur oxidation, we conducted a microcosm-based pilot study with tailings of an active pond. Incubations were performed under oxic and anoxic conditions, with and without KNO3 as an electron acceptor and thiosulfate as a common substrate for microbial sulfur oxidation. The highest potentials of sulfur oxidation occurred in oxic assays (1.21 mmol L−1 day−1). Under anoxic conditions, rates were significantly lower and dominated by chemical transformation (0.09 mmol L−1 day−1; p < 0.0001). The addition of KNO3 to anoxic incubations increased microbial thiosulfate oxidation 2.5-fold (0.23 mmol L−1 day−1; p = 0.0474), with complete transformation to SO42− coupled to NO3− consumption, pointing to the activity of sulfur-oxidizing bacteria (SOB) under nitrate-reducing conditions. Importantly, in the presence of KNO3, a decrease in sedimentary sulfides was associated with an increase in S0, which indicates the potential for microbially mediated oxidation of sulfide minerals and ARD generation. Furthermore, the comparative analysis of sediments from other anthropogenic aquatic habitats demonstrated high similarities with respect to viable SOB counts and corresponding activity rates.

Transcriptome Analysis of Environmental Pseudomonas Isolates Reveals Mechanisms of Biodegradation of Naphthenic Acid Fraction Compounds (NAFCs) in Oil Sands Tailings

Naphthenic acid fraction compounds (NAFCs) are highly recalcitrant constituents of oil sands tailings. Although some microorganisms in the tailings can individually and synergistically metabolize NAFCs, the biochemical mechanisms that underpin these processes are hitherto unknown. To this end, we isolated two microorganisms, Pseudomonas protegens and Pseudomonas putida, from oils sands tailings and analyzed their transcriptomes to shed light on the metabolic processes employed by them to degrade and detoxify NAFCs. We identified 1048, 521 and 1434 genes that are upregulated in P. protegens, P. putida and a 1:1 co-culture of the strains, respectively. We subsequently enumerated the biochemical activities of enriched genes and gene products to reveal the identities of the enzymes that are associated with NAFC degradation. Separately, we analyzed the NAFCs that are degraded by the two pseudomonads and their 1:1 co-culture and determined the composition of the molecules using mass spectrometry. We then compared these molecular formulas to those of the cognate substrates of the enriched enzymes to chart the metabolic network and understand the mechanisms of degradation that are employed by the microbial cultures. Not only does the consortium behave differently than the pure cultures, but our analysis also revealed the mechanisms responsible for accelerated rate of degradation of NAFCs by the co-culture. Our findings provide new directions for engineering or evolving microorganisms and their consortia for degrading NAFCs more stably and aggressively.

Methanogenic Biodegradation of iso-Alkanes by Indigenous Microbes from Two Different Oil Sands Tailings Ponds

iso-Alkanes, a major fraction of the solvents used in bitumen extraction from oil sand ores, are slow to biodegrade in anaerobic tailings ponds. We investigated methanogenic biodegradation of iso-alkane mixtures comprising either three (2-methylbutane, 2-methylpentane, 3-methylpentane) or five (2-methylbutane, 2-methylpentane, 2-methylhexane, 2-methylheptane, 2-methyloctane) iso-alkanes representing paraffinic and naphtha solvents, respectively. Mature fine tailings (MFT) collected from two tailings ponds, having different residual solvents (paraffinic solvent in Canadian Natural Upgrading Limited (CNUL) and naphtha in Canadian Natural Resources Limited (CNRL)), were amended separately with the two mixtures and incubated in microcosms for ~1600 d. The indigenous microbes in CNUL MFT produced methane from the three-iso-alkane mixture after a lag of ~200 d, completely depleting 2-methylpentane while partially depleting 2-methylbutane and 3-methylpentane. CNRL MFT exhibited a similar degradation pattern for the three iso-alkanes after a lag phase of ~700 d, but required 1200 d before beginning to produce methane from the five-iso-alkane mixture, preferentially depleting components in the order of decreasing carbon chain length. Peptococcaceae members were key iso-alkane-degraders in both CNUL and CNRL MFT but were associated with different archaeal partners. Co-dominance of acetoclastic (Methanosaeta) and hydrogenotrophic (Methanolinea and Methanoregula) methanogens was observed in CNUL MFT during biodegradation of three-iso-alkanes whereas CNRL MFT was enriched in Methanoregula during biodegradation of three-iso-alkanes and in Methanosaeta with five-iso-alkanes. This study highlights the different responses of indigenous methanogenic microbial communities in different oil sands tailings ponds to iso-alkanes.